| 牛瑟氏泰勒虫HSP70基因的克隆及原核表达 |
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| 引用本文： | 金超,贾立军,薛书江,王娜,钱年超,张守发.牛瑟氏泰勒虫HSP70基因的克隆及原核表达[J].黑龙江畜牧兽医,2010(11). |
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| 作者姓名： | 金超 贾立军 薛书江 王娜 钱年超 张守发 |
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| 作者单位： | 延边大学农学院动物医学系; |
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| 基金项目： | 国家自然科学基金项目(30560112) |
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| 摘 要： | 根据GenBank(D12692.1)上发表的牛瑟氏泰勒虫HSP70基因序列设计、合成1对特异性的引物,采用PCR方法扩增牛瑟氏泰勒虫HSP70基因片段,将扩增产物与pMD18-Tsimple载体连接,重组质粒经PCR、酶切鉴定后测序;用pGEX-4T-1表达载体构建重组质粒pGEX-HSP70,经IPTG诱导表达后进行SDS-PAGE、Western-blot分析。结果表明:扩增的HSP70基因与D12692.1序列同源性为99.3%;表达的融合蛋白分子质量约为68ku,而且可与牛瑟氏泰勒虫阳性血清发生特异性反应。
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| 关 键 词： | 牛瑟氏泰勒虫 HSP70 克隆 原核表达 |
Cloning and procaryotic expression of HSP70 gene of bovine Theileria sergenti |
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| JIN Chao,JIA Li-jun,XUE Shu-jiang,WANG Na,QIAN Nian-chao,ZHANG Shou-fa.Cloning and procaryotic expression of HSP70 gene of bovine Theileria sergenti[J].Heilongjiang Animal Science And veterinary Medicine,2010(11). |
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| Authors: | JIN Chao JIA Li-jun XUE Shu-jiang WANG Na QIAN Nian-chao ZHANG Shou-fa |
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| Institution: | JIN Chao,JIA Li-jun,XUE Shu-jiang,WANG Na,QIAN Nian-chao,ZHANG Shou-fa(Department of Veterinary Medicine,Agricultural College of Yanbian University,Longjing 133400,China) |
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| Abstract: | A pair of specific primers were designed and synthetized according to the HSP70 gene sequence(D12692.1)of bovine Theileria sergenti which was published in GenBank.HSP70 gene fragment of bovine Theileria sergenti was amplified by PCR and ligated into pMD18-T simple vector.The recombinant plasmid was sequenced after being certified by PCR amplification and restriction analysis.The pGEX-HSP70 was constructed using pGEX-4T-1 and analyzed by SDS-PAGE and Western-blot after it was induced and expressed by IPTG.Th... |
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| Keywords: | Bovine Theileria sergenti HSP70 clone procaryotic expression |
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